Performing Department
(N/A)
Non Technical Summary
Salish Kootenai College (SKC) on the Flathead Reservation in Montana houses the Life Sciences Department and their laboratories for novel research of interest to the Selis, Ksanka, and Qlispepeople. This new discovery grant application is designed to investigate the impact of bison on the Montana grasslands, with the Bison Range on the Flathead Reservation as the key study site. Objectives are to investigate the biogeochemical cycling of the soil with bison dung and to identify the plants eaten by the bison using DNA sequence analysis from dung samples. The overreaching hypothesis is that bison, who consume native plants are aiding the natural grassland environment. Undergraduate research at SKC will examine these objectives increasing the institutional capacity by training students at a Tribal college on laboratory techniques and to increase the body of knowledge for the Tribal management of the Bison Range.
Animal Health Component
25%
Research Effort Categories
Basic
75%
Applied
25%
Developmental
0%
Goals / Objectives
The major goalof this project isto understand the role of bison on grasslands as they are reintroduced throughout Montana including Tribal lands. Due to climate change, there is an increased interest in carbon capture by soil and soil health through native plant growth and recycling.Recent efforts have been made to reintroduce bison populations across the American west, mainly led by tribal nations. Bison have been reintroduced across portions of the vast Northern Great Plains (NGP), which is the largest intact grassland region in North America covering over 180 million acres according to the USDA Natural Resources Conservation Service. These bison introductions offer a unique opportunity to study the reintroduction of a large herbivore on an intact grassland. Bison are a key spiritual and historically relevant animals, it is also important to recognize the reintroduction of bison to Montana grasslands as culturally significant to Tribal stakeholders throughout the state. Other reservations, such as Fort Belknap and the Blackfeet reservation are in the process of bison re-introductions. Providing scientifically sound research activities to measure soil and native plant health in bison herd ranges, is important in herd monitoring in relationship to the grasslands and climate change cycles. Objectives of this project are (1) connect the bison to grazing of native plants of Montana grasslands and (2) connect the natural roaming bison to the soil nutrient cyclins in grasslands.Measurable outcomes from objective (1) DNA plant sequences for identification from bison dung identifying plant species. Measurable outcomes from objective (2) Soil moisture measurements, β-glucosidase activity measurements, and total carbon measurements from soil underneath bison dung.
Project Methods
Efforts:DNA Extraction: Scatt and dung samples will be stored in the Life Sciences Department at -80? until sample preparation for DNA extraction. DNA extraction will be carried out using the PureLink™ Microbiome DNA Purification Kit from ThermoFisher (Cat. A29790). Sample procedure is for the purification of high-quality host DNA from stool samples pub. no. MAN0014334 rev. A.0. The kit provides spin-column technology for purified DNA ready for downstream PCR of the internal transcribed spacer region of nuclear ribosomal DNA (ITS).PCR amplification products will be gel purified and quality samples will be used for library sequencing.In-House Sequencing: After library preparation and quality confirmed, samples will be sequenced on the iSeq™ 100 Illumina sequencer in the Life Sciences labs at SKC.Evaluation:Sequence Analysis: The iSeq™ 100 has phasing correction for base correction if a base read falls behind or jumps ahead and quality scoring (Q-score) to calculate error probabilities with the higher Q-score as a lower error probability. These measures are reported in real time for every cycle through the run. Output data is directly linked to the cloud-sourced BaseSpace Sequence Hub for which SKC Life Sciences has a current and ongoing subscription.Efforts:Five soil cores will be taken at one site and homogenized into a single sample.The activity of three microbial enzymes - β-glucosidase, peroxidase, and urease - will be analyzed using colorimetric spectrophotometry.The soil suspension will be aliquoted and prepared according to the procedure in the β-glucosidase Assay Kit (catalog number ab272521) from Abcam.The soil suspension will be aliquoted and prepared according to the procedure in the Urease Activity Assay Kit (colorimetric) (catalog number ab204697) from Abcam.200 g subsamples of soil will be sent to the Oregon State University (OSU) Soil Health Lab to be analyzed for total carbon and soil respiration (a proxy for microbial biomass).Evaluation:Statistical Analysis: Means of the bison grazed samples and control samples for each response variable will be compared using a t-test. Means and 95% confidence intervals will be reported for each response variable. Bison grazed and control samples will be compared using a general linear mixed-effects model (GLM) where grazing is a fixed-effect and site-identity is a random-effect. This will account for variation between site locations. An individual GLM will be reported for each variable, in addition to a conditional R2 that represents the influence from fixed-effects and site-specific variability. Statistical analysis will be done in R 4.2.2 (R Core Team, 2022).